SUMMARY Combining DNA-demethylating agents (DNA methyltransferase inhibitors [DNMTis]) with histone deacetylase inhibitors (HDACis) holds promise for enhancing cancer immune therapy. Herein, pharmacologic and isoform specificity of HDACis are investigated to guide their addition to a DNMTi, thus devising a new, low-dose, sequential regimen that imparts a robust anti-tumor effect for non-small-cell lung cancer (NSCLC). Using in-vitro-treated NSCLC cell lines, we elucidate an interferon α/β-based transcriptional program with accompanying upregulation of antigen presentation machinery, mediated in part through double-stranded RNA (dsRNA) induction. This is accompanied by suppression of MYC signaling and an increase in the T cell chemoattractant CCL5. Use of this combination treatment schema in mouse models of NSCLC reverses tumor immune evasion and modulates T cell exhaustion state towards memory and effector T cell phenotypes. Key correlative science metrics emerge for an upcoming clinical trial, testing enhancement of immune checkpoint therapy for NSCLC.

The liver is the most common site of metastatic disease1. While this metastatic tropism may reflect mechanical trapping of circulating tumor cells, liver metastasis is also dependent, at least in part, on formation of a “pro-metastatic” niche that supports tumor cell spread to the liver2,3. Mechanisms that direct formation of this niche, though, are poorly understood. Here, we show that hepatocytes coordinate myeloid cell accumulation and fibrosis within the liver, and in doing so, increase the susceptibility of the liver to metastatic seeding and outgrowth. Early during pancreatic tumorigenesis, hepatocytes demonstrate activation of Signal Transducer and Activator of Transcription 3 (STAT3) signaling and increased production of serum amyloid A1 and A2 (SAA). Overexpression of SAA by hepatocytes also occurs in pancreatic and colorectal cancer patients with liver metastases, and many patients with locally advanced and metastatic disease display elevated levels of circulating SAA. STAT3 activation in hepatocytes and the subsequent production of SAA are dependent on interleukin 6 (IL-6) that is released into the circulation by non-malignant cells. Genetic ablation or blockade of components of IL-6/STAT3/SAA signaling prevents establishment of a pro-metastatic niche and inhibits liver metastasis. Our data reveal an intercellular network underpinned by hepatocytes that forms the basis for a pro-metastatic niche in the liver and identify new therapeutic targets.

This paper is meant to be an introduction to and general reference for ultrasound imaging for new and moderately experienced users of the instrument. The paper consists of eight sections. The first explains how ultrasound works, including beam properties, scan types and machine features. The second section discusses image quality, including the interpretation of anatomical features and artefacts seen in the image. The third section discusses the validity of the data collection procedures, including the effects of stabilizing the transducer and head position, and discusses some methods for stabilization. Section four discusses validation of the ultrasound and stabilization systems. The fifth section presents a sample recording set-up, supplemental information, and normalization strategies for sessions and subjects. In section six are methods of extracting contours from ultrasound images, displaying them, and analysing them. The seventh section considers the tracking of pellets on the tongue surface and the differences between tracking tissue points and continuous surfaces. The last section presents methods, challenges and results of 3D, computerized reconstruction of tongue surfaces. An outline of the paper can be found in Appendix I.

Ovarian cancer is the most lethal of all gynecological cancers, and there is an urgent unmet need to develop new therapies. Epithelial ovarian cancer (EOC) is characterized by an immune suppressive microenvironment, and response of ovarian cancers to immune therapies has thus far been disappointing. We now find, in a mouse model of EOC, that clinically relevant doses of DNA methyltransferase and histone deacetylase inhibitors (DNMTi and HDACi, respectively) reduce the immune suppressive microenvironment through type I IFN signaling and improve response to immune checkpoint therapy. These data indicate that the type I IFN response is required for effective in vivo antitumorigenic actions of the DNMTi 5-azacytidine (AZA). Through type I IFN signaling, AZA increases the numbers of CD45 immune cells and the percentage of active CD8 T and natural killer (NK) cells in the tumor microenvironment, while reducing tumor burden and extending survival. AZA also increases viral defense gene expression in both tumor and immune cells, and reduces the percentage of macrophages and myeloid-derived suppressor cells in the tumor microenvironment. The addition of an HDACi to AZA enhances the modulation of the immune microenvironment, specifically increasing T and NK cell activation and reducing macrophages over AZA treatment alone, while further increasing the survival of the mice. Finally, a triple combination of DNMTi/HDACi plus the immune checkpoint inhibitor α-PD-1 provides the best antitumor effect and longest overall survival, and may be an attractive candidate for future clinical trials in ovarian cancer.

This paper presents three-dimensional tongue surfaces reconstructed from multiple coronal cross-sectional slices of the tongue. Surfaces were reconstructed for sustained vocalizations of the American English sounds [symbol: see text]. Electropalatography (EPG) data were also collected for the sounds to compare tongue surface shapes with tongue-palate contact patterns. The study was interested also in whether 3-D surface shapes of the tongue were different for consonants and vowels. Previous research and speculation had found that there were differences in production, acoustics, and linguistic usage between the two groups. The present study found that four classes of tongue shape were adequate to categorize all the sounds measured. These classes were front raising, complete groove, back raising, and two-point displacement. The first and third classes have been documented before in the midsagittal plane [cf. R. Harshman, P. Ladefoged, and L. Goldstein, J. Acoust. Soc. Am. 62, 693-707 (1976)]. The first three classes contained both vowels and consonants, the last only consonants. Electropalatographic patterns of the sounds indicated three categories of tongue-palate contact: bilateral, cross-sectional, and combination of the two. Vowels used only the first pattern, consonants used all three. The EPG data provided an observable distinction in contact pattern between consonants and vowels. The ultrasound tongue surface data did not. The conclusion was that the tongue actually has a limited repertoire of shapes and positions them against the palate in different ways for consonants versus vowels to create narrow channels, divert airflow, and produce sound.

Using noninvasive real-time ultrasound, tongue movement was visualized during single swallowing in eight normal subjects and one neurologically impaired patient with dysphagia and chronic aspiration. In normals, a clearly defined muscular wave of the tongue, traveling at approximately 15 cm/sec, carried a 5-cc test water bolus posteriorly. In the patient who had 12th cranial nerve weakness, there was complete absence of normal tongue activity and no midtongue bolus formation or transmission.

Abstract. Six female adults were studied during the production of four single swallows each, using real-time ultrasound. A pellet was affixed to the tongue at the junction between the (calculated) anterior third and posterior two-thirds of the tongue surface. Direction, rate, and extent of pellet movement were measured and used to create stages of tongue blade movement during swallowing. Pellet movement was then compared to the three stages of hyoid movement (ascent, steady, descent). Anterior-posterior and superior-inferior components of pellet movement were examined and discussed.

Point-tracking techniques provide timing information about structural movements of the tongue. Imaging techniques provide information about cross-sectional and pharyngeal tongue shape and movement. This study joined these techniques in a single subject. Five pellets on the tongue surface were tracked using x-ray microbeam, and the midsagittal and coronal planes of the tongue were imaged using real-time ultrasound. The speech materials were the consonants [s] and [l] and the vowels [i], [a], and [o] combined in VCVCe utterances. Analyses concentrated on the difference in tongue movements related to the two consonants. A model of tongue movement was developed, in which critical features of consonant shape and position dominated the tongue opening movement. In this model, the tongue is divided into subdivisions termed "functional segments" in both the sagittal and coronal planes. Movements of the functional segments created observable opening movement patterns.